Analyzing DNA barcoding and identifying toxins caused by neurotoxic mushroom poisoning using liquid chromatography tandem mass spectrometry

• Published in: Journal of associated medical sciences Vol. 57; no. 2; pp. 107 - 114
• Main Authors: Phatsarapongkul, Sriprapa, Parnmen, Sittiporn, Nooron, Nattakarn, Chankunasuka, Rungsaeng, Thunkhamrak, Chidkamon, Nitma, Unchalee, Palakul, Nisakorn, Chonnakijkul, Pornpanna, Sikaphan, Sujitra, Uttawichai, Chutimon, Polputpisatkul, Dutsadee, Rojanawiwat, Archawin
• Format: Journal Article
• Language: English
• Published: 04.05.2024
• ISSN: 2539-6056; 2539-6056
• DOI: 10.12982/JAMS.2024.032

Background: Neurotoxic mushroom poisoning often exhibits rapid symptom onset, typically attributed to compounds such as Ibotenic acid, which affect the central nervous system. This study addresses a new case of mushroom-related food poisoning in southern Thailand. Objective: The objectives are to determine the presence of ibotenic acid in cases of mushroom-related food poisoning utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and to identify toxic Amanita species implicated in these cases. Materials and methods: Remnant mushroom samples obtained from three clinically reported cases were used. Nucleotide similarity was compared against the rRNA/ ITS databases using NCBI BLAST search. Phylogenetic analyses were conducted using maximum likelihood (ML) and FastTree approaches. LC-MS/MS was employed to separate of Ibotenic acid, determine its molecular weight and perform precursor ion fragmentation. Results: Analysis of the rRNA/ITS databases revealed a high nucleotide similarity between suspected mushroom samples and Amanita digitosa. Detailed phylogenetic analysis confirmed that mushroom samples from the three poisoning cases clustered with A. digitosa. LC-MS/MS analysis showed the presence of ibotenic acid, with precursor ion (m/z 159) and product ion (m/z 113.1) as the major toxic substances. Clinically, patients poisoned by ibotenic acid-containing mushrooms exhibited a short latent period with symptoms of nausea, vomiting, vertigo, delirium, confusion, and fatigue. Conclusion: The genus Amanita comprises both edible and inedible species that produce several lethal toxins. The report of ibotenic acid in A. digitosa is a novel finding, valuable for food safety monitoring and healthcare decision-maker. This is especially notable due to the accuracy and rapidity of the analytical process.